Difference between revisions of "NMDA Receptors"

From MagnetoWiki
Jump to navigation Jump to search
(→‎Primers for Davenport's Real-Time PCR experiment: added new primers designed in May 2009)
(cleaned up)
Line 2: Line 2:
  
 
Primers were ordered 2009 May from Sigma Aldrich
 
Primers were ordered 2009 May from Sigma Aldrich
 +
 +
(designed using [http://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi PrimerPlus3])
 +
  
 
<tab class="wikitable" border = "1" cellspacing="3" cellpadding = "3" head = top>
 
<tab class="wikitable" border = "1" cellspacing="3" cellpadding = "3" head = top>
Line 31: Line 34:
 
=Primers for Real-Time PCR=
 
=Primers for Real-Time PCR=
  
Taken from Yamamoto et al. 2008 PMID 17957211 using Applied Biosystems
+
Looked at primers from Yamamoto et al. 2008 PMID 17957211 using Applied Biosystems, but ended up designing own primers above.
 
 
 
 
NR1
 
Forward primer 5'-GTTCTTCCGCTCAGGCTTTG-3'
 
Reverse primer 5'-AGGGAAACGTTCTGCTTCCA-3'
 
TaqMan probe 5'-FAM-CGGCATGCGCAAGGACAGCC-TAMRA-3'
 
 
NR2A
 
Forward primer 5'-AGCCCCCTTCGTCATCGTA-3'
 
Reverse primer 5'-GACAGGGCACCGTGTTCCT-3'
 
TaqMan probe 5'-FAM-AGGACATAGACCCCCTGACTGAGACCTGTG-TAMRA-3'
 
 
NR2B
 
Forward primer 5'-CCCCCAAGTTCTGGTTGGT-3'
 
Reverse primer 5'-TTTTGGGAACGAGCTTTGCT-3'
 
TaqMan probe 5'-FAM-TTGGCCGTCTTGGCCGTATCAGG-TAMRA-3'
 
 
NR2C
 
Forward primer 5'-GGGTGATGATGTTCGTGATGTG-3'
 
Reverse primer 5'-GTGAGGTTCTGGTTGTAGCTGACA-3'
 
TaqMan probe 5'-FAM-TCACGGTGGTTGCCATCACTGTCTTC-TAMRA-3'
 
 
 
 
 
 
 
 
 
Primers chosen for experiment (according to Andre's advice)
 
 
 
(designed using [http://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi PrimerPlus3]
 
 
 
 
 
 
 
NR1  U11418.1
 
2360 Forward ttcacagaagtgcgatctgg, Tm 60.0
 
2467 Reverse atggacagggaaacgttctg, Tm 60.0
 
Product size of 108 bp
 
 
 
 
 
Serine Racemase    NM_198757
 
 
 
856 forward: tggtagatgcgttggtggta, Tm 60.0
 
958  reverse: tcagcagcgtacaccttcac, Tm60.1
 
 
 
674-807, product size 134bp
 
 
Rat GAPDH
 
 
 
Forward TCACTGCCACTCAGAAGACTG
 
Reverse GGATGACCTTGCCCACAGC
 
  
  
  
 
[[Category:Taste_Aversion]]
 
[[Category:Taste_Aversion]]

Revision as of 16:07, 23 May 2009

Primers for Davenport's Real-Time PCR experiment

Primers were ordered 2009 May from Sigma Aldrich

(designed using PrimerPlus3)


Probe Abbr Sequence Tm Size Genbank Reference
Rat NR1 subunit QNR1FOR ttcacagaagtgcgatctgg 60.0 108 U11418.1,2360-2467 PrimerPlus
QNR1REV atggacagggaaacgttctg 60.0
Rat Serine Racemase QSERRACFOR tggtagatgcgttggtggta 60.0 103 NM_198757, 856-958 PrimerPlus
QSERRACREV tcagcagcgtacaccttcac 60.1
Rat GADPH QGADPHFOR tcactgccactcagaagactg 60 ??? Andre designed with sequencher
QGADPHREV ggatgaccttgcccacagc 60 . . .

Primers were ordered 2009 March from Sigma Aldrich (NB: these primers worked fine in a test RT-PCR, but Andre said the product sizes should be 100-150bp, and the Tm's should be closer together. So, we redesigned and ordered primers above).

Probe Abbr Sequence Tm Size Genbank Reference
NR1 RTNR1FOR GTTCTTCCGCTCAGGCTTTG 66.0 66 RNU11418, 2396-2461 Yamamoto et al. 2008 PMID 17957211
RTNR1REV AGGGAAACGTTCTGCTTCCA 65.8
Serine Racemase RTSERRACFOR ATTGCAAGAAACTGGCCATC 63.9 295 NM198757, 664-958 Yoshikawa et al., 2006 PMID 16973158
RTSERRACREV TCAGCAGCGTACACCTTCAC 64.1
GADPH RTGADPHFOR TGCACCACCAACTGCTTAG 62.5 201 DQ403053.1, 397-597 Uno et al., 2002 PMID 12068078
RTGADPHREV GGATGCAGGGATGATGTTC 63.0
Note: GADPHFOR and REV maybe mixed up . . . .

Primers for Real-Time PCR

Looked at primers from Yamamoto et al. 2008 PMID 17957211 using Applied Biosystems, but ended up designing own primers above.